Fluid handling in mesoscale analytical devices

• US 5,304,487 A
• Filed: 05/01/1992
• Issued: 04/19/1994
• Est. Priority Date: 05/01/1992
• Status: Expired due to Term

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Kricka, Larry J., Wilding, Peter, Zemel, Jay N.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Torres, Ramon
• Application Number:
  US07/877,536
• Time in Patent Office: 718 Days
• Field of Search: 422/55, 422/58, 422/61, 436/524, 436/164, 436/807, 436/809, 436/49, 436/501, 436/180, 435/2, 435/7.21, 435/359, 435/291, 435/6, 435/7.2
• US Class Current: 435/29
• CPC Class Codes: B01F 25/41 Mixers of the fractal type
• CPC Class Codes: B01F 35/7182 with means for feeding the material with a fractal or tree-type distribution in a surface
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: Y10S 366/03 Micromixers: variable geometry from the pathway influences mixing/agitation of non-laminar fluid flow
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

Devices are provided for analyzing a fluid cell containing sample. The devices comprise a solid substrate, microfabricated to define at least one sample inlet port and a mesoscale flow system. The mesoscale flow system includes a sample flow channel, extending from the inlet port, and a cell handling region for treating cells disposed in fluid communication with the flow channel. The devices may further include a structure inducing flow of cells in the sample through the flow system. In one embodiment, the cell-handling region may comprise a cell lysis structure to enable the lysis of cells in the sample, prior to, e.g., the detection of an intracellular component in the cell sample. In another embodiment, the cell handling region may comprise a cell capture region, comprising binding sites which reversibly bind to a specific population of cells in the cell sample, to permit the isolation of the specific cell population from the sample. The devices can be utilized in a wide range of automated sensitive and rapid tests for the analysis of a fluid cell containing sample.

Claims

1. A device for analyzing a fluid, cell-containing sample, the device comprising:

a solid substrate microfabricated to define;

a sample inlet port; and

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

a cell handling region for treating cells disposed in fluid communication with said flow channel, said cell handling region comprising a cell lysing structure; and

means for detecting an analyte in a fluid sample in said flow system.

2. The device of claim 1 further comprising means for inducing flow of cells in a sample through said mesoscale flow channel and said cell handling region to force cells in said sample into contact with said cell lysing structure, thereby to lyse cells in said sample.

3. The device of claim 1 wherein said cell lysing structure comprises a portion of a flow channel having cell membrane piercing protrusions extending from a wall thereof.

4. The device of claim 1 wherein said cell lysing structure comprises sharp edged particles trapped within said cell handling region;

and

wherein said device further comprises means for inducing flow to force cells in said sample into contact with said sharp edged particles, thereby to lyse said cells.

5. The device of claim 1 wherein said cell lysing structure comprises a region of restricted cross-sectional dimension sufficient to permit passage of intracellular molecules while prohibiting passage of cells.

6. The device of claim 1 wherein said means for detecting comprises means downstream of said cell lysing structure for detecting the presence of an intracellular molecular component of a cell in said sample.

7. The device of claim 1 further comprising means disposed downstream of said cell lysing structure for collecting insoluble cellular debris.

8. The device of claim 1 further comprising a filter means disposed downstream of said cell lysing structure.

9. The device of claim 1, wherein said substrate comprises microfabricated silicon.

17. The device of claim 1, 10 or 15, wherein said means for detecting comprises an appliance for use in combination with said substrate, said appliance comprising:

means for holding said substrate; and

optical means for viewing the contents of said mesoscale flow system in said substrate.

18. The device of claim 17, wherein said optical means comprises magnifying optics and a video camera, and wherein said appliance further comprises:

a tilt mechanism for manually adjusting the angle and location of the device; and

a video screen for viewing the contents of said flow system.

24. The device of claim 1, 10, 15 or 23 wherein, within at least a portion of a channel in a said flow system, the channel width and channel depth each are between 0.1 μ

m and 500 μ

m.

25. The device of claim 24 wherein the channel width in said portion is between 2.0 and 500 μ

m.

26. The device of claim 24 wherein the channel depth in said portion is between 0.1 and 100 μ

m.

10. A device for analyzing a fluid, cell-containing sample, the device comprising:

a solid substrate microfabricated to define;

a sample inlet port; and

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

a cell handling region for treating cells disposed in fluid communication with said flow channel, said cell handling region comprising a cell capture region comprising immobilized binding sites which reversibly bind a preselected cell surface molecule of a cell population in a cell-containing fluid sample; and

means for inducing flow of cells in said sample through said mesoscale flow channel and said cell handling region;

wherein said means for inducing flow of said cell-containing sample is used to induce flow;

at a first flow rate sufficiently slow to permit capture of cells in said cell population by said binding sites, thereby to separate said cell population from said sample; and

at a second flow rate, higher than said first flow rate, and sufficient to release said separated cells from said capture region; and

wherein said device further comprises means for detecting an analyte in a fluid sample in said flow system.

11. The device of claim 10 wherein said detecting means comprises means downstream of said cell capture region for determining the presence of an extracellular component of said sample.

12. The device of claim 10 wherein said flow system further comprises:

cell lysing means downstream from said cell capture region and wherein said flow inducing means includes means for forcing cells into said cell lysing means, thereby to lyse cells in said sample; and

wherein said detecting means comprises means for detecting the presence of an intracellular component in said captured cells.

13. The device of claim 12 further comprising filter means, disposed between said cell-lysing means and said detection means, for filtering cellular debris from said sample.

14. The device of claim 13 further comprising a sump for collecting insoluble debris disposed adjacent said filter.

16. The device of claim 2, 10 or 15 further comprising an appliance for use in combination with said substrate, said appliance comprising:

means for holding said substrate; and

fluid input means interfitting with an inlet port on said substrate; and

wherein said means for inducing flow comprises pump means, disposed in said appliance, for passing fluid through the flow system of said substrate when it is held in said housing means.

19. The device of claim 2, 10 or 15, wherein said:

a solid substrate microfabricated to define;

a sample inlet port;

a mesoscale flow system further comprises;

a branching channel in fluid communication with said flow channel; and

at least two additional ports communicating between said flow channel and said branching channel, respectively, and the exterior of said flow system; and

wherein said device further comprises valve means for directing flow through said flow system to a selected one of said additional ports.

20. The device of claim 19 wherein said detecting means comprises a detection region within said mesoscale flow system for optically or electrically gathering data indicative of the presence or concentration of an analyte in a sample contained within said flow system.

21. The device of claim 19 further comprising an appliance for use in combination with said substrate, said appliance comprising:

means for holding said substrate;

fluid flow channels interfitting with at least two of said ports when said substrate is held in said holding means; and

wherein said means for inducing flow comprises pump means disposed within said appliance in fluid communication with said inlet ports for inducing flow within said flow system.

22. The device of claim 21 wherein said valve means is disposed within said appliance.

15. A device for analyzing a fluid, cell-containing sample, the device comprising:

a solid substrate microfabricated to define;

a sample inlet port; and

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

a cell handling region for treating cells disposed in fluid communication with said flow channel, said cell handling structure defining;

a cell sieve comprising means defining a plurality of flow passages of restricted size allowing only cells of a sufficiently small diameter to pass therethrough; and

a cell lysing structure;

means for inducing flow of cells in a sample through said mesoscale flow channel and said cell handling structure; and

means for detecting an analyte in a fluid sample in said flow system.

23. A device for analyzing a cell-containing fluid sample, the device comprising:

a solid substrate microfabricated to define;

a mesoscale cell handling structure; and

at least two mesoscale flow systems, each of which comprise a flow channel and an analyte detection region, one of said flow systems being adapted to analyze a sample, the other being adapted as a control, and said flow systems being in fluid communication with said cell handling structure; and

means for inducing flow of a sample through said cell handling structure and then through both said flow systems, thereby to permit comparison of data from the detection regions of said systems.